Preparation of 2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamide hydrochloride and other derivatives of 2-hydroxy-1,3-diamines

ABSTRACT

The present invention provides a new process for the preparation of 2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamide hydrochloride, wherein this compound is prepared directly from the chloromethylalcohol. Importantly, the process of the present invention results in higher yields of 2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamide hydrochloride without sacrificing its purity. The processes of the present invention can be used to prepare not only the 2R,3S-derivative, but also the 2S,3S-, 2S,2R- and the 2R,3R-derivatives.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a division of application Ser. No. 12/190,713, filedAug. 13, 2008 (now U.S. Pat. No. 7,870,850, issued Oct. 5, 2010), whichis a division of application Ser. No. 11/052,934, filed Feb. 7, 2005(now U.S. Pat. No. 7,414,444, issued Aug. 19, 2008), which is a divisionof application Ser. No. 10/382,185, filed Mar. 4, 2003 (now U.S. Pat.No. 6,852,887, issued Feb. 8, 2005), which is a division of applicationSer. No. 09/747,372, filed Dec. 22, 2000 (now U.S. Pat. No. 6,548,706,issued Apr. 15, 2003), which claims the benefit of U.S. ProvisionalPatent Application No. 60/172,043, filed Dec. 23, 1999, which areincorporated herein by reference in their entirety for all purposes.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

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BACKGROUND OF THE INVENTION

Human immunodeficiency virus (HIV), the causative agent of acquiredimmunodeficiency syndrome (AIDS), encodes three enzymes, including thewell-characterized proteinase belonging to the aspartic proteinasefamily, the HIV protease. Inhibition of this enzyme has been regarded asa promising approach for treating AIDS. Hydroxyethylamine isosteres havebeen extensively utilized in the synthesis of potent and selective HIVprotease inhibitors. However, this modern generation of HIV proteaseinhibitors has created an interesting challenge for the syntheticorganic chemist. Advanced x-ray structural analysis has allowed for thedesign of molecules that fit closely into active sites on enzymescreating very effective drug molecules. Unfortunately, these molecules,designed by molecular shape, are often difficult to produce usingconventional chemistry.

The modern generation of HIV inhibitors has structural similarities in acentral three-carbon piece containing two chiral carbons that link twolarger groups on each side (see, e.g., Parkes, et al., J. Org. Chem.,39:3656 (1994)). In general, the chemical bond from the central part toone of the larger groups is a carbon-nitrogen bond which is usuallyaccomplished by reacting an epoxide with an amine.2S,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride is a key intermediate in the synthesis of proteaseinhibitors (see, e.g., U.S. Pat. Nos. 5,585,397, 5,723,490 and 5,783,701as well as PCT International Publication No. WO 94/0563, the teachingsof all of which are incorporated herein by reference). The processcurrently used to prepare2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride is illustrated in FIG. 1.

As illustrated in FIG. 1,2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride is prepared by a 4-step process starting from acommercially available (Aerojet Fine Chemicals (Sacramento, Calif.))starting material, 2S,3S-chloromethylalcohol (2S,3S-CMA). In the firststep, 2S,3S-CMA is reacted with sodium hydroxide in THF/ethanol to givethe corresponding epoxide in 91% yield. In the second step, the epoxideis dissolved in toluene and reacted with excess isobutylamine at atemperature of about 75° C. to about 80° C. to give Compound I. Reactionof Compound I with p-nitrobenzenesulfonyl chloride (i.e., nosylchloride) in toluene at a temperature of about 85° C. to about 90° C.,followed by deprotecting of the Boc protecting group with aqueous HCl ata temperature of about 85° C. to about 90° C. gives Compound III, i.e.,2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride, in 64% overall yield.

Unfortunately, this process has a number of drawbacks. Some of thedrawbacks associated with this process include high reactiontemperatures which lead to the formation of side products; lowsolubility of intermediates in the reaction solvent which requires highprocessing temperatures during work-up which, in turn, leads to longercycle times, material handling losses, etc.; and the like. As such,there remains a need in the art for an improved process for preparing2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride which overcomes the drawbacks associated with thecurrently used process. Quite surprisingly, the present inventionfulfills this and other needs.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a new process for the preparation of2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride, i.e., Compound III. In this new process, one of thechemical steps (i.e., the epoxide step) as well as material handlinglosses associated with the previously used process have been eliminated.More particularly, using the process of the present invention, thehalomethylalcohol, i.e., HMA, (e.g., chloromethylalcohol) issuccessfully converted to Compound III directly without the need toisolate the Boc-epoxide. Numerous publications have described thesynthesis of Compound I-like materials, but they all involve reaction ofan epoxide with an amine. As such, the process of the present inventionis the first example of a process wherein a protease inhibitorintermediate is prepared directly from the halomethylalcohol. Moreover,the process of the present invention results in a higher yield ofCompound III, while not sacrificing its purity. In addition, the processof the present invention allows for the use of the lower purity HMA,thereby eliminating a purification step. Finally, it is pointed out thatelimination of the epoxide step eliminates the need to isolate a toxicintermediate (i.e., a mutagen) and, thus, circumvents safety issuesinvolved with dust explosivity of the Boc-epoxide.

Other features, objects and advantages of the invention and itspreferred embodiments will become apparent from the detailed descriptionwhich follows.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates the process previously used to prepare2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride.

DETAILED DESCRIPTION OF THE INVENTION

It has now been discovered that a protease inhibitor intermediate, i.e.,2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride, can be prepared directly from a halomethylalcohol (HMA)(e.g., chloromethylalcohol) without having to isolate the Boc-epoxide.As such, the present invention provides a new process for thepreparation of2R,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride, i.e., Compound III, wherein this compound is prepareddirectly from a HMA. Importantly, the process of the present inventionresults in higher yields of Compound III, while not sacrificing itspurity. Moreover, the process of the present invention can be used toprepare not only the 2S,3S-derivative, but also the 2R,3S-, 2S,2R- andthe 2R,3R-derivatives.

As such, the first step in the process for preparing a Compound ofFormula III involves the preparation of a compound having the followinggeneral formula:

the process comprising: combining a halomethylalcohol, e.g.,chloromethylalcohol, with an amine, e.g., isobutylamine (IBA), to form areaction mixture, and heating the reaction mixture until the reaction iscomplete, thereby forming the compound of Formula I, wherein the HMA hasthe following general formula:

In the above process, X is a leaving group (e.g., a halo group, such aschloro, bromo, fluoro or iodo, a tosylate group, a mesylate group,etc.); R¹ is an amino acid side chain or a functionalized amino acidside chain (e.g., an alkyl group, a benzyl group, a phenyl thiomethylgroup, a protected hydroxymethyl group, an S-alkyl group, anitrobenzenyl group, etc.); R² is a protecting group (e.g., t-Boc(tert-butyloxycarboxy), i-Boc (iso-butyloxycarboxy), Cbz(benzyloxycarboxy), MOC (methoxycarboxy), Fmoc(fluorenylmethoxycarbonyl), dibenzyl, etc.); and R³ and R⁴ areindependently selected and are functional groups including, but notlimited to, hydrogen, alkyl, cycloalkyl, aryl and —NHP, wherein P isselected from the group consisting of H, alkyl, cycloalkyl, aryl and aprotecting group or, alternatively, R³ and R⁴ together can form a cyclicstructure including, but not limited to, quinoline and isoquinoline,etc.

In a preferred embodiment, the HMA is a 2S,3S-halomethylalcohol(2S,3S-HMA). In a presently preferred embodiment, the 2S,3S-HMA has thefollowing formula:

In another preferred embodiment, the Compound of Formula I has thefollowing formula:

In the above process, a HMA is reacted with an amine at a temperatureranging from about 50° C. to 100° C. to give the compound of Formula Iin high yields. In a presently preferred embodiment, the HMA is reactedwith an amine at a temperature ranging from about 60° C. to about 75° C.to give the Compound of Formula I in very high yields (about 98-99%yield). The amine used in the above process can be a primary amine, asecondary amine, a substituted amine (e.g., N-substituted sulfonamides,etc.), hydrazines and substituted hydrazines (e.g., N-substitutedhydrazine, etc.). Exemplar amines include, but are not limited to,methylamine, ethylamine, isopropylamine, n-butylamine, isobutylamine,cyclohexylamine, benzylamine, paranitrobenzylamine, dimethylamine,isobutylmethylamine, decahyroquinoline, decahydroisoquinoline,pyrimidine, isobutylparanitrobenzenesulfonamide,methylparanitrobenzenesulfonamide, isobutylmetanitrobenzenesulfonamide,hydrazine, substituted hydrazines, etc.

In a presently preferred embodiment, the amine employed in the aboveprocess is isobutylamine (IBA). Importantly, the solvent employed in thepreviously used method (i.e., toluene) has been eliminated and, in thisembodiment, the reaction is conducted in neat IBA, using IBA as thereaction solvent. This simplifies the IBA recovery process (nofractional distillation/column needed to separate IBA from toluene), andallows for the reaction to be carried out at slightly lowertemperatures. It has been found, however, that under theses conditions,the reaction times can be relatively long (˜30 h). To reduce reactiontimes, it has been discovered that an acid scavenger can be added to thereaction mixture.

As such, in one embodiment of the above process, an acid scavenger isadded to the reaction mixture to reduce reaction times. In a preferredembodiment, the acid scavenger has the general formula: M⁺B⁻, wherein M⁺is selected from the group consisting of K⁺, Na⁺, Li⁺, Ca⁺², Mg⁺² andAl⁺²; and B⁻ is selected from the group consisting of ⁻OH, HCO₃ ⁻ andCO₃ ⁻². In a preferred embodiment, the acid scavenger is sodiumbicarbonate. In another preferred embodiment, the acid scavenger ispotassium bicarbonate. When an acid scavenger is added to the reactionmixture, about 0.5 to 2 equivalents are added. In a presently preferredembodiment, about 1 equivalent of the acid scavenger is added to thereaction mixture. It has been found that by adding even only 1.05equivalents of an acid scavenger (e.g., sodium bicarbonate) to thereaction mixture, the reaction times can be reduced from 30 hours toless than 4 hours. Moreover, this reduction in reaction times can beachieved without sacrificing product quality and/or yield. It will bereadily apparent to those of skill in the art that other acidscavengers, base phosphates, etc. can be used in the methods of thepresent invention to reduce reaction times.

Thus, as illustrated in Table I, the reaction rates of the basecatalyzed reaction of chloromethylalcohol (CMA) with isobutylamine arecomparable to those of the epoxide with isobutylamine, with the reactionbeing substantially complete in 3 hours.

TABLE I Rxn Time (h) % Compound I 0 0.15 1 90.8 2 97.7 3 98.6 4 98.8

Moreover, it is pointed out that the Boc-epoxide employed in thepreviously used method is thermally unstable, and at higher temperatures(>70° C.) will undergo a deprotecting reaction to give free amine which,in turn, leads to a variety of side-reactions. In contrast, HMA, e.g.,CMA, is more thermally stable than Boc-epoxide. The higher thermalstability of HMA, e.g., CMA, coupled with lower reaction/processingtemperatures allows for Compound III to be prepared in a much higheryield and purity. Moreover, using the above process, the purity ofisolated Compound I is in the neighborhood of 98.7 to 98.8% (area %) byHPLC. Typical results for the reaction of 2S,3S-CMA with isobutylamineto give Compound I are illustrated below:

TABLE II Reaction # Scale* Yield Purity CPO86-135 (HP) 100 g 98.5% 98.8%CPO86-143 (HP) 100 g 98.5% 98.7% CPO86-147 (HP) 100 g ND 98.8% CPO86-160(HP) 250 g   99% 98.8%

In one embodiment, the above process further comprises recovering (e.g.,isolating and purifying) the Compound of Formula I from the reactionmixture. However, in a presently preferred embodiment, the Compound ofFormula I is filtered and used in the next step without first beingpurified.

The second step in the process for preparing a Compound of Formula IIIinvolves the preparation of a compound having the following generalformula:

the process comprising: (a) combining p-nitrobenzenesulfonyl chloride,i.e., nosyl chloride, with an alkyl acetate to form a first reactionmixture; (b) combining a tertiary amine (e.g., triethylamine (TEA)) witha compound of Formula I in an aromatic solvent (e.g., toluene) to form asecond reaction mixture, the compound of Formula I having the followinggeneral formula:

and, (c) adding the first reaction mixture with the second reactionmixture to form a third reaction mixture, and heating the third reactionmixture until the reaction is complete, thereby forming the compound ofFormula II.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; and R³ and R⁴ areindependently selected and are functional groups including, but notlimited to, hydrogen, alkyl, cycloalkyl, aryl and —NHP, wherein P isselected from the group consisting of H, alkyl, cycloalkyl, aryl and aprotecting group or, alternatively, R³ and R⁴ together can form a cyclicstructure including, but not limited to, quinoline and isoquinoline. Ina presently preferred embodiment, the compound of Formula I has thefollowing general formula:

In another preferred embodiment, the Compound of Formula II has thefollowing general formula:

In the first step of the above process, nosyl chloride is combined withan alkyl acetate to form a first reaction mixture. Alkyl acetatessuitable for use include, but are not limited to, methyl acetate, ethylacetate, isopropyl acetate, isobutyl acetate, etc. In a presentlypreferred embodiment, ethyl acetate is employed. In the second step ofthe above process, a tertiary amine is combined with a Compound ofFormula I in an aromatic solvent to form a second reaction mixture.Exemplar tertiary amines include, but are not limited to, triethylamine,diisopropylethylamine, pyridine and N-methylmorpholine. In a presentlypreferred embodiment, triethylamine is used. Suitable aromatic solvents,include, but are not limited to, toluene, benzene and xylene. In apresently preferred embodiment, toluene is used. In the third step ofthe above process, the first reaction mixture is added to the secondreaction mixture to form a third reaction mixture, and the thirdreaction mixture is heated until the reaction is complete. In apresently preferred embodiment, the first reaction mixture is heated orwarmed prior to adding it to the second reaction mixture. The thirdreaction mixture is heated to a temperature ranging from about 50° C. toabout 100° C. and, more preferably, to a temperature ranging from about60° C. to about 75° C.

Ethyl acetate is a good solvent for Compound I and Compound IIintermediates; however, under certain conditions, Compound I will reactwith ethyl acetate to give N-acetyl Compound I. As such, it has now beensurprisingly discovered that the best way to carry out the aboveprocess, i.e., the second step in the preparation of the Compound ofFormula III, is to dissolve the nosyl chloride in ethyl acetate (nosylchloride is not very soluble in toluene), and then add it to a solutionof the Compound of Formula I in toluene that has been warmed to atemperature of about 70° C. Under these conditions, the reaction isinstantaneous and is complete when the addition is done. Moreover, sincenosyl chloride is much more reactive than ethyl acetate, formation ofthe N-acetyl Compound I is not detected. Further, addition of ethylacetate to the reaction mixture improves the solubility of Compound IIin the reaction mixture, thereby allowing the processing to be conductedat about 70° C. instead of at about 85° C. Use of ethyl acetate as aco-solvent also improves the partitioning of the Compound of Formula IIinto the organic phase and, thus, minimizes loss due to product loss inaqueous phase. Further, use of lower reaction temperature (70° C. versus85-90° C.) also improves the quality of the Compound of Formula II sinceside reactions, such as de-protecting, etc., are minimized at lowertemperatures.

In an alternative embodiment, the Compound of Formula II can be prepareddirectly from a HMA. As such, the present invention provides a processfor the preparation of a compound having the general formula:

the process comprising: combining a halomethyl alcohol (HMA) with asulfonamide to form a reaction mixture, and heating said reactionmixture until the reaction is complete, thereby forming the compound ofFormula II, wherein said HMA has the following general formula:

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; R³ is a functionalgroup including, but not limited to, hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H, alkyl,cycloalkyl, aryl and a protecting group; R⁴ is a functional groupincluding, but not limited to, alkyl, cycloalkyl and aryl; and X is aleaving group.

In a preferred embodiment, the HMA is a 2S,3S-halomethyl-alcohol(2S,3S-HMA). In a presently preferred embodiment, the 2S,3S-HMA has thefollowing formula:

In the above process, the HMA is reacted with a sulfonamide. Suitablesulfonamides include those having the following general formula:

wherein: R⁵ and R⁶ are independently selected and are functional groupsincluding, but not limited to, alkyl, cycloalkyl and aryl. In apreferred embodiment, R⁵ is alkyl and R⁶ is aryl. In a further preferredembodiment, the aryl has the general formula:

wherein: Z is a functional group including, but not limited to,hydrogen, alkyl, halogen, hydroxy, alkoxy, —NO₂ and —NAP, wherein P is aprotecting group. In a preferred embodiment, the sulfonamide is selectedfrom the group consisting of isobutylparanitrobenzenesulfonamide,isobutylmetanitrobenzenesulfonamide andisobutylorthonitrobenzenesulfonamide.

In one embodiment of the above process, an acid scavenger can be addedto the reaction mixture to reduce reaction times. In a preferredembodiment, the acid scavenger has the general formula: M⁺B⁻, wherein M⁺is selected from the group consisting of K⁺, Na⁺, Li⁺, Ca⁺², Mg⁺² andAl⁺²; and B⁻ is selected from the group consisting of ⁻OH, HCO₃ ⁻ andCO₃ ⁻². In a preferred embodiment, the acid scavenger is sodiumbicarbonate. In another preferred embodiment, the acid scavenger ispotassium bicarbonate. When an acid scavenger is added to the reactionmixture, about 0.5 to 2 equivalents are added. In a presently preferredembodiment, about 1 equivalent of the acid scavenger is added to thereaction mixture. In this process, a HMA is reacted with a sulfonamideat a temperature ranging from about 50° C. to 125° C. to give thecompound of Formula II in high yields. In a presently preferredembodiment, the HMA is reacted with a sulfonamide at a temperatureranging from about 60° C. to about 80° C. to give the Compound ofFormula II in very high yields.

The final step in the process of the present invention involves theconversion of a Compound of Formula II to a Compound of Formula III, theCompound of Formula III having the following general formula:

in which R¹ is an amino acid side chain or a functionalized amino acidside chain; R³ is a functional group including, but not limited tohydrogen, alkyl, cycloalkyl, aryl and —NHP, wherein P is selected fromthe group consisting of H, alkyl, cycloalkyl, aryl and a protectinggroup; and X⁻ is the conjugate base of an acid having a pK_(a) of lessthan about 4. The process for converting a Compound of Formula II to aCompound of Formula III comprises: (a) combining a Compound of FormulaII with an aromatic solvent/alkyl acetate (e.g., toluene/ethyl acetate)to form a reaction mixture, and heating the reaction mixture, theCompound of Formula II having the following general formula:

and, (b) treating the reaction mixture with an acid having a pK_(a) ofless than about 4 (e.g., hydrochloric acid (HCl)) until the reaction iscomplete, thereby forming the Compound of Formula III. In a presentlypreferred embodiment, this process further comprises recovering theCompound of Formula III from the reaction mixture by cooling to 5° C.and filtering.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; and R³ is a memberselected from the group consisting of hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H, alkyl,cycloalkyl, aryl and a protecting group; and X⁻ is the conjugate base ofsaid acid.

In a presently preferred embodiment the Compound of Formula II has thefollowing formula:

In another preferred embodiment, the Compound of Formula III has thefollowing general formula:

In step (b) of the above process, the reaction mixture is treated orcontacted with an acid having a pK_(a) of less than about 4. Suitableacids include, but are not limited to, HCl, HBr, CF₃CO₂H, CF₃SO₃H,CH₃SO₃H, H₂SO₄, citric acid, tartaric acid, oxalic acid and p-toluenesulfonic acid. The reaction mixture is treated with the acid for a timeperiod of at lease about 0.5 hours and, typically, for a period of timeranging from about 1 to about 5 hours. In a presently preferredembodiment, the acid is aqueous hydrochloric acid (HCl). In thisembodiment, the hydrochloric acid can be, for example, about 37% aqueoushydrochloric acid. In another preferred embodiment, the hydrochloricacid is gaseous hydrochloric acid. In this embodiment, the gaseoushydrochloric acid can be bubbled through the reaction mixture. Inanother presently preferred embodiment, the acid is methanesulfonic acid(CH₃SO₃H). In a presently preferred embodiment, the Compound of FormulaIII is recovered from the reaction mixture.

In addition to the foregoing, it has also been discovered that ahalogenated solvent (e.g., methylene chloride) is a superior solventthan toluene or toluene/ethyl acetate for synthesizing the Compounds ofFormulae II and III. The higher solubility of Compound I-IIIintermediates in a halogenated solvent (e.g., methylene chloride) allowsfor the reactions to be carried out at much lower temperatures (about 30to 35° C. lower than the toluene/ethyl acetate process and about 45-50°C. lower than the toluene process). The ability to conduct the reactionsat lower temperatures has a profound effect on the quality and yield ofthe resulting material. Low reaction temperatures minimize decompositionand other side-reactions (e.g., deprotecting), thereby leading toproducts with fewer impurities which, in turn, leads to improved qualityand higher yield (15 to 20% higher yield). It has been found that therate of the deprotecting reaction with, for example, aqueous HCl is muchslower in a halogenated solvent (e.g., methylene chloride) than intoluene or toluene/ethyl acetate. For example, in toluene/ethyl acetatethe time required to achieve >99% conversion is about 3 hours, whereasin methylene chloride, it is nearly 5 days. However, it has surprisinglybeen discovered that the use of, for example, gaseous HCl, instead ofaqueous HCl, will reduce the reaction time from 5 days to 3 hours. Anadditional advantage for using gaseous HCl is that it does not requireadditional stripping to remove the water. When HCl is employed as theacid, the final product (Compound III) is a hydrochloride salt, and itis important to remove water in order to achieve a good yield. However,this problem is eliminated when anhydrous gaseous HCl is used. Moreover,the product quality of crude Compound III prepared by the halogenatedsolvent route is much better than that of crude Compound III prepared bythe toluene or toluene/ethyl acetate processes. Consequently, the yieldof the isolated Compound III is nearly 15% higher with the halogenatedsolvent process. In addition, the intermediates do not have to beisolated when a halogenated solvent (e.g., methylene chloride) is used.It is noted that, e.g., the sulfonamide impurity is soluble in methylenechloride; otherwise, it is very difficult to remove.

As such, in one embodiment, the present invention provides a process forpreparing a compound having the following general formula:

the process comprising: (a) combining nosyl chloride with a halogenatedsolvent (e.g., methylene chloride) to form a first reaction mixture; (b)combining a tertiary amine (e.g., triethylamine) with a compound ofFormula I in a halogenated solvent (e.g., methylene chloride) to form asecond reaction mixture, the compound of Formula I having the followinggeneral formula:

and, (c) adding the first reaction mixture to the second reactionmixture to form a third reaction mixture, and heating the third reactionmixture to a temperature ranging from about 10° C. to about 50° C. and,more preferably, to a temperature ranging from about 30° C. to about 35°C. until the reaction is complete, thereby forming the compound ofFormula II. In a presently preferred embodiment, the first reactionmixture is heated or warmed prior to adding it to the second reactionmixture.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; and R³ is a functionalgroup including, but not limited to, hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H, alkyl,cycloalkyl, aryl and a protecting group.

In a presently preferred embodiment, the Compound of Formula I has thefollowing formula:

In another preferred embodiment, the Compound of Formula II has thefollowing formula:

In the above process, a halogenated solvent is employed. Suitablehalogenated solvents include, but are not limited to, methylenechloride, chloroform, trifluorotoluene (oxsol),parachlorotrifluorotoluene and trichloroethylene. In a preferredembodiment, the halogenated solvent is methylene chloride. Suitabletertiary amines include, but are not limited to, triethylamine,diisopropylethylamine, pyridine and N-methylmorpholine. In a preferredembodiment, the tertiary amine is triethylamine.

In another embodiment, the present invention provides a process forpreparing a compound having the following general formula:

the process comprising: (a) combining a compound of Formula II with ahalogenated solvent (e.g., methylene chloride) to form a reactionmixture and heating the reaction mixture to a temperature ranging fromabout 10° C. to about 50° C. and, more preferably, to a temperatureranging from about 30° C. to about 35° C., the compound of Formula IIhaving the following general formula:

and, (b) treating the reaction mixture with an acid having a pK_(a) ofless than about 4 (e.g., hydrochloric acid) until the reaction iscomplete, thereby forming the compound of Formula III. Exemplarhalogenated solvents includes, but are not limited to, the following:methylene chloride, chloroform, trifluorotoluene (oxsol),parachlorotrifluorotoluene and tricloroethylene. In a presentlypreferred embodiment, the halogenated solvent is methylene chloride.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; and R³ is a memberselected from the group consisting of hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H alkyl,cycloalkyl, aryl and a protecting group; and X⁻ is the conjugate base ofthe acid employed in the reaction.

In a presently preferred embodiment, the Compound of Formula II has thefollowing formula:

In another preferred embodiment, the Compound of Formula III has thefollowing general formula:

In step (b) of the above process, the reaction mixture is treated orcontacted with an acid having a pK_(a) of less than about 4. Exemplaracids include, but are not limited to, HCl, HBr, CF₃CO₂H, CF₃SO₃H,CH₃SO₃H, H₂SO₄, citric acid, tartaric acid, oxalic acid and p-toluenesulfonic acid. The reaction mixture is treated with the acid until thereaction is complete and, typically, for a time period of at lease about0.5 hours and, more preferably, for a period of time ranging from about1 to about 5 hours. In one preferred embodiment, the hydrochloric acidis aqueous hydrochloric acid. In this embodiment, the hydrochloric acidcan be, for example, about 37% aqueous hydrochloric acid. In anotherpreferred embodiment, the hydrochloric acid is gaseous hydrochloricacid. In this embodiment, the gaseous hydrochloric acid can be bubbledthrough the reaction mixture. In another preferred embodiment, the acidis methanesulfonic acid (CH₃SO₃H). In a presently preferred embodiment,the Compound of Formula III is recovered from the reaction mixture.

In another embodiment, the present invention provides a process forpreparing a Compound of Formula III from a HMA, the Compound of FormulaIII having the following general formula:

the process comprising: (a) combining a halomethyl alcohol (HMA) and anamine (e.g., isobutylamine (IBA)) in a halogenated solvent (e.g.,methylene chloride) with an acid scavenger to form a reaction mixture,and heating the reaction mixture to a temperature ranging from about 10°C. to about 50° C. and, more preferably, to a temperature ranging fromabout 30° C. to about 35° C., wherein the HMA has the following generalformula:

(b) adding to the first reaction mixture nosyl chloride while stirringthe reaction mixture, and recovering the organic layer of the reactionmixture to form a solution; and (c) reacting the solution with an acidhaving a pKa of less than about 4 until the reaction is complete,thereby forming the compound of Formula III.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; R³ is a functionalgroup including, but not limited to, hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H, alkyl,cycloalkyl, aryl and a protecting group; and L is a leaving group.

In a presently preferred embodiment, the HMA is a 2S,3S-HMA. In apresently preferred embodiment, the 2S,3S-HMA has the following formula:

In another presently preferred embodiment, the Compound of Formula IIIhas the following formula:

In the above process, an amine is combined with a halogenated solvent.Suitable amines include, but are not limited to, primary amines,secondary amines, substituted amines, hydrazines and substitutedhydrazines. Exemplar amines include, but are not limited to,methylamine, ethylamine, isopropylamine, n-butylamine, isobutylamine,cyclohexylamine, benzylamine, paranitrobenzylamine, dimethylamine,isobutylmethylamine, decahyroquinoline, decahydroisoquinoline,pyrimidine, isobutylhydrazine and t-Boc-hydrazine. In a preferredembodiment, the amine is a primary amine. In a presently preferredembodiment, the amine is isobutylamine (IBA), preferably neat IBA. Inthe above process, a halogenated solvent is employed. Exemplarhalogenated solvents includes, but are not limited to, the following:methylene chloride, chloroform, trifluorotoluene (oxsol),parachlorotrifluorotoluene and trichloroethylene. In a presentlypreferred embodiment, the halogenated solvent is methylene chloride.

In step (c) of the above process, the solution from step (b) is treatedor contacted with an acid having a pK_(a) of less than about 4. Exemplaracids include, but are not limited to, HCl, HBr, CF₃CO₂H, CF₃SO₃H,CH₃SO₃H, H₂SO₄, citric acid, tartaric acid, oxalic acid and p-toluenesulfonic acid. The solution is treated with the acid until the reactionis complete. Typically, the solution is treated for a time period of atlease about 0.5 hours and, more preferably, for a time period rangingfrom about 1 to about 5 hours. In one preferred embodiment, the acid ishydrochloric acid. In another preferred embodiment, the acid ismethanesulfonic acid (CH₃SO₃H).

In yet another embodiment, the present invention provides a process forpreparing a Compound of Formula III from a compound of Formula I, theCompound of Formula III having the following general formula:

the process comprising: (a) adding a Compound of Formula I in ahalogenated solvent (e.g., methylene chloride) with a tertiary amine andnosyl chloride to form a reaction mixture, and heating the reactionmixture at a temperature ranging from about 10° C. to about 50° C. and,more preferably, to a temperature ranging from about 30° C. to about 35°C.; the compound of Formula I having the following general formula:

and (b) reacting the reaction mixture with an acid having a pK_(a) ofless than about 4 until the reaction is complete, thereby forming theCompound of Formula III.

In the above process, R¹ is an amino acid side chain or a functionalizedamino acid side chain; R² is a protecting group; and R³ is a memberselected from the group consisting of hydrogen, alkyl, cycloalkyl, aryland —NHP, wherein P is selected from the group consisting of H alkyl,cycloalkyl, aryl and a protecting group; and X⁻ is the conjugate base ofthe acid used in step (b).

In a presently preferred embodiment, the Compound of Formula I has thefollowing formula:

In another preferred embodiment, the Compound of Formula III has thefollowing formula:

In the above process, a tertiary amine is combined with a halogenatedsolvent. Suitable tertiary amines include, but are not limited to,triethylamine, diisopropylethylamine, pyridine and N-methylmorpholine.In a presently preferred embodiment, triethylamine is used. Exemplarhalogenated solvents includes, but are not limited to, the following:methylene chloride, chloroform, trifluorotoluene (oxsol),parachlorotrifluorotoluene and trichloroethylene. In a presentlypreferred embodiment, the halogenated solvent is methylene chloride.

In step (b) of the above process, the reaction mixture is reacted withan acid having a pK_(a) of less than about 4. Exemplar acids include,but are not limited to, HCl, HBr, CF₃CO₂H, CF₃SO₃H, CH₃SO₃H, H₂SO₄,citric acid, tartaric acid, oxalic acid and p-toluene sulfonic acid. Thesolution is treated with the acid until the reaction is complete.Typically, the solution is treated for a time period of at least about0.5 hours and, more preferably, for a time period ranging from about 1to about 5 hours. In one preferred embodiment, the acid is hydrochloricacid. In another preferred embodiment, the acid is methanesulfonic acid(CH₃SO₃H).

In a presently preferred embodiment, the process further compriseswashing the reaction mixture; heating the reaction mixture to atemperature ranging from about 40° C. to about 45° C.; and thenazeotropically drying the reaction mixture prior to step (b). In anotherpreferred embodiment, the Compound of Formula III is recovered from thereaction mixture. In one such embodiment, the recovery step comprises:(i) cooling the reaction mixture to a temperature of about −5° C.; (ii)filtering the reaction mixture to remove theN-isobutyl-p-nitrobenzenesulfonamide impurities and provide the compoundof Formula III; and (iii) washing the compound of Formula III and thendrying it in vacuo.

Using the above processes,2S,3S-N-isobutyl-N-(2-hydroxy-3-amino-4-phenylbutyl)-p-nitrobenzenesulfonylamidehydrochloride can be prepared directly from the chloromethylalcohol(CMA) without having to isolate the Boc-epoxide. The process of thepresent invention is the first demonstration of a process wherein aprotease inhibitor intermediate is prepared directly from thehalomethylalcohol. Again, this is extremely helpful because eliminationof the epoxide eliminates the need to isolate a toxic intermediate(i.e., a mutagen) and, thus, circumvents safety issues involved withdust explosivity of the Boc-epoxide. Importantly, the process of thepresent invention results in higher yields of Compound III, while notsacrificing its purity.

Those of skill in the art will readily understand that the term “alkyl,”as used herein, refers to a branched or unbranched, saturated orunsaturated, monovalent hydrocarbon radical having from 1-12 carbonsand, preferably, from 1-6 carbons. When the alkyl group has from 1-6carbon atoms, it is referred to as a “lower alkyl.” Suitable alkylradicals include, for example, methyl, ethyl, n-propyl, i-propyl,2-propenyl (or allyl), n-butyl, t-butyl, i-butyl (or 2-methylpropyl),etc. As used herein, the term alkyl encompasses “substituted alkyls.”Substituted alkyl refers to alkyl as just described including one ormore functional groups such as lower alkyl, aryl, aralkyl, acyl, halogen(i.e., alkylhalos, e.g., CF₃), hydroxy (e.g., hydroxymethyl), amino,alkylamino, acylamino, acyloxy, alkoxy (e.g., methoxymethyl), mercaptoand the like. These groups may be attached to any carbon atom of thelower alkyl moiety.

The term “cycloalkyl,” by itself or in combination with other terms,represents cyclic versions of “alkyl.” Examples of cycloalkyl includecyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl,and the like. The cycloalkyl group may be substituted or unsubstituted.

The term “alkoxy” is used herein to refer to the —OR group, where R is alower alkyl, substituted lower alkyl, aryl, substituted aryl, aralkyl orsubstituted aralkyl. Suitable alkoxy radicals include, for example,methoxy, ethoxy, phenoxy, t-butoxy (e.g., methoxyethoxy, methoxymethoxy,etc.), etc.

The term “aryl,” as used herein, refers to an aromatic substituent whichmay be a single ring or multiple rings which are fused together, linkedcovalently, or linked to a common group such as an ethylene or methylenemoiety. The aromatic ring(s) may include phenyl, naphthyl, biphenyl,diphenylmethyl, 2,2-diphenyl-1-ethyl, and may contain a heteroatom, suchas thienyl, pyridyl and quinoxalyl. The aryl group may also besubstituted with halogen atoms, or other groups such as nitro, carboxyl,alkoxy, phenoxy, and the like. Additionally, the aryl group may beattached to other moieties at any position on the aryl radical whichwould otherwise be occupied by a hydrogen atom (such as 2-pyridyl,3-pyridyl and 4-pyridyl).

In view of the foregoing, it will be readily apparent to those of skillin the art that the foregoing processes can be used to prepare not onlythe 2S,3S-derivative, but also the 2R,3S-, 2S,2R- and the2R,3R-derivatives. Again, the process of the present invention resultsin higher yields of these derivatives, while not sacrificing theirpurity.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of noncriticalparameters which can be changed or modified to yield essentially thesame results.

EXAMPLES I. Process A: Toluene/Ethyl Acetate Process

A. Preparation of Compound I

A mixture of 2S,3S-CMA (251.8 g, 0.84 mol), isobutylamine 737.8 g (10.1mol) and sodium hydrogen carbonate (77.6 g, 92 mol) in water (323.9 g)is heated at a gentle reflux (˜60° C.) for 5 h. Water (1000 g) is addedand excess isobutyl amine is removed by distillation under nitrogen atan internal reaction temperature of 70° C. (The recovery isapproximately 85% and the recovered IBA can be recycled). Additionalwater (500 g) is added and the product is isolated by filtration. Theprecipitated solid is washed with water and dissolved in toluene (1500g). The solution is reduced in volume by ⅓ by distillation at 70° C.under reduced pressure (300 mm-Hg) to remove water and residualisobutylamine. The solution of Compound I in toluene is used directly insynthesis of Compound II. However, in order to obtain yield/purity data,Compound I is isolated by evaporation of the solvent and the resultingsolid is dried in oven to constant weight. The yields normally range inthe neighborhood of 98.5 to 99% with purities (HPLC) ranging in theneighborhood of 98.7 to 98.8%. The isolated product was characterized by1H/¹³C NMR and by HPLC/TLC analysis.

B. Preparation of Compound II

Solution of Compound I in toluene was warmed to 70° C. and treated withtriethylamine (220 g, 2.18 mol). Then, a solution ofp-nitrobenzenesulfonyl chloride (195 g, 0.88 mol) in ethyl acetate (1 l)was added over 3 h. The organic mixture was then washed sequentially at65-70° C. with water (400 mL), 5% aqueous sodium hydrogen carbonate (400mL) and water (400 mL). The product is obtained as a solution intoluene/ethyl acetate and is used directly in synthesis of Compound III.The yields, as determined by evaporating the solvent and isolating crudeCompound II, were in the neighborhood of 97 to 98%.

C. Preparation of Compound III

The solution of Compound II in toluene/ethyl acetate at 65° C.-70° C.was treated with 37% aqueous hydrochloric acid (160 g, 1.64 mol) over aperiod of 2 h. The mixture was then distilled at 70° C. under reducedpressure (500-600 mm-Hg) until the residual ethyl acetate content wasless than 5 mol % in toluene (as determined by ¹H NMR analysis), and thewater content was less than 1% (as determined by Karl Fischer analysis).When distillation was complete, the toluene volume was adjusted to 15%solids and ethanol (360 mL, denatured with 5 wt % Methanol) was added.The mixture was cooled to ambient temperature over 2 h, held at ambientfor 1 h, then cooled to −5° C. to −10° C. for 1 h. The precipitatedsolid was isolated by filtration, washed with toluene and n-hexanes, anddried in vacuo at 80° C. to give 247 g (73%) of Compound III. The purityof Compound III as determined by HPLC was 99.2 wt % (99.8 area %).Compound III was also characterized by 1H/13C NMR analysis.

II. Process B: Methylene Chloride Process

A. One Pot Process

A solution of 2S,3S-CMA (44 g, 0.146 mol), isobutylamine (11 g, 0.150mol) in 270 mL of methylene chloride was stirred with sodium hydrogencarbonate (33 g, 0.4 mol) in water (100 g) for 5 h. is heated at agentle reflux (˜30° C.) for 6 h. Then nosyl chloride (35.4 g, 0.157moles) and triethylamine (17.7 g, 0.175 moles) was added and the mixturestirred for 6 h at 30-35° C. The organic layer was removed and theresidue washed with water (100 mL). Gaseous HCl was bubbled through thesolution for 2-3 hours until the reaction was complete as indicated byHPLC analysis. The residual acid is removed with a stream of nitrogengas and the reaction mixture was cooled to −5° C. for 1 h. The mixturewas filtered to give 60 g (89%) of Compound III with 99.7% purity byHPLC.

B. Process Starting with Compound I

A solution of triethylamine (17.7 g, 0.175 mol) and Compound I (49.0 g1.46 mol) in methylene chloride (270 mL) was heated to 30-35° C. andnosyl chloride (35.4 g, 0.157 mol) in methylene chloride was added over3 h. The solution is heated to 35° C. and washed sequentially with water(65 mL), 5% sodium bicarbonate solution (60 mL) and water (80 mL). Thesolution was heated reflux and water removed azeotropically. When KarlFischer analysis of the solution indicated that less than 0.1% of waterremained, the solution was cooled to ambient. Hydrogen chloride gas waspassed through the solution for 3 h until HPLC analysis indicated thatthe reaction was complete. The mixture was cooled to 0-5° C. andfiltered to give a white solid. The solid was washed with hexanes (65mL) and dried in vacuo to give 59.5 g (89%) of compound III that was99.7% pure by HPLC analysis.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference for allpurposes.

What is claimed is:
 1. A process for preparing a compound having thefollowing general formula:

said process comprising: (a) combining a halomethyl alcohol (HMA) and anamine in a halogenated solvent with aqueous bicarbonate to form areaction mixture, and heating said reaction mixture to a temperatureranging from about 10° C. to about 50° C., wherein said HMA has thefollowing general formula:

(b) adding to said first reaction mixture nosyl chloride while stirringsaid reaction mixture, and recovering the organic layer of said reactionmixture to form a solution; and (c) reacting said solution with an acidhaving a pK_(a) of less than about 4 until the reaction is complete,thereby forming the compound of Formula III; wherein: R¹ is an aminoacid side chain or a functionalized amino acid side chain; R² is aprotecting group; R³ is a member selected from the group consisting ofhydrogen, alkyl, cycloalkyl, aryl and —NHP, wherein P is selected fromthe group consisting of H, alkyl, cycloalkyl, aryl and a protectinggroup; X⁻ is the conjugate base of said acid; and L is a leaving group.2. The process in accordance with claim 1, wherein said amine is amember selected from the group consisting of primary amines, secondaryamines, substituted amines, hydrazines and substituted hydrazines. 3.The process in accordance with claim 1, wherein said amine is a memberselected from the group consisting of methylamine, ethylamine,isopropylamine, n-butylamine, isobutylamine, cyclohexylamine,benzylamine, paranitrobenzylamine, dimethylamine, isobutylmethylamine,decahyroquinoline, decahydroisoquinoline, pyrimidine, isobutylhydrazineand t-Boc-hydrazine.
 4. The process in accordance with claim 3, whereinsaid amine is isobutylamine (IBA).
 5. The process in accordance withclaim 1, wherein said halogenated solvent is a member selected from thegroup consisting of methylene chloride, chloroform, trifluorotoluene(oxsol), parachlorotrifluorotoluene and trichloroethylene.
 6. Theprocess in accordance with claim 1, wherein said halogenated solvent ismethylene chloride.
 7. The process in accordance with claim 1, whereinsaid acid is a member selected from the group consisting of HCl, HBr,CF₃CO₂H, CF₃SO₃H, CH₃SO₃H, H₂SO₄, citric acid, tartaric acid, oxalicacid and p-toluene sulfonic acid.
 8. The process in accordance withclaim 7, wherein said acid is HCl.
 9. The process in accordance withclaim 1, wherein said HMA is a 2S,3S-HMA.
 10. The process in accordancewith claim 9, wherein said 2S,3S-HMA has the following general formula:


11. The process in accordance with claim 10, wherein said compound ofFormula III has the following general formula:


12. The process in accordance with claim 1, wherein said reactionmixture is heated to a temperature ranging from about 30° C. to about35° C.
 13. A process for preparing a compound having the followinggeneral formula:

said process comprising: (a) adding a compound of Formula I in ahalogenated solvent with a tertiary amine and nosyl chloride to form areaction mixture, and heating said reaction mixture at a temperatureranging from about 10° C. to about 50° C., said compound of Formula Ihaving the following general formula:

(b) reacting said reaction mixture with an acid having a pK_(a) of lessthan about 4 until the reaction is complete, thereby forming thecompound of Formula III; wherein: R¹ is an amino acid side chain or afunctionalized amino acid side chain; R² is a reacting group; R³ and R⁴are independently selected from the group consisting of hydrogen, alkyl,cycloalkyl, aryl and —NHP, wherein P is selected from the groupconsisting of H, alkyl, cycloalkyl, aryl and a protecting group, or R³and R⁴ together form a cyclic structure selected from the groupconsisting of quinoline and isoquinoline; and X⁻ is the conjugate baseof said acid.
 14. The process in accordance with claim 13, wherein saidcompound of Formula I has the following general formula:


15. The process in accordance with claim 14, wherein said compound ofFormula III has the following general formula:


16. The process in accordance with claim 13, wherein said halogenatedsolvent is a member selected from the group consisting of methylenechloride, chloroform, trifluorotoluene (oxsol),parachlorotrifluorotoluene and trichloroethylene.
 17. The process inaccordance with claim 13, wherein said halogenated solvent is methylenechloride.
 18. The process in accordance with claim 13, wherein said acidis a member selected from the group consisting of HCl, HBr, CF₃CO₂H,CF₃SO₃H, CH₃SO₃H, H₂SO₄, citric acid, tartaric acid, oxalic acid andp-toluene sulfonic acid.
 19. The process in accordance with claim 18,wherein said acid is HCl.
 20. The process in accordance with claim 13,further comprising washing, heating and azeotropically drying saidreaction mixture prior to step (b).
 21. The process in accordance withclaim 13, further comprising: (c) recovering said compound of FormulaIII from said reaction mixture.
 22. The process in accordance with claim13, wherein said reaction mixture is heated to a temperature rangingfrom about 30° C. to about 35° C.